Determination of amikacin in body fluid by high-performance liquid-chromatography with chemiluminescence detection

A simple and sensitive method was developed for the quantification of amikacin in human plasma and urine samples. The method involves centrifugation of body fluid plasma after dilution with an ethanol/sodium carbonate mixture, and then an aliquot of the supernatant is directly injected into the chromatograph. After separation on a reversed-phase C18 column (runtime 20 min), aminoglycoside is detected on the basis of its complex formation reaction with Cu(II), the catalyst of the lummol/hydrogen peroxide chemiluminescence system. Using a volume of 500 mu l biological sample, linearity is established over the concentration range 0.15-2.0 mu g/ml and the limit of detection (LOD) is ca. 50 mu g/l in plasma or urine. The intra-day and inter-day precision (measured by relative standard deviation, R.S.D.%) are always less than 9%, and relative recoveries are found to be over 92%. (c) 2006 Elsevier B.V. All rights reserved.